Characterization of a novel androgen receptor mutation in a relapsed CWR22 prostate cancer xenograft and cell line

Clifford G Tepper, David L. Boucher, Philip E. Ryan, Ai Hong Ma, Liang Xia, Li Fen Lee, Thomas G. Pretlow, Hsing-Jien Kung

Research output: Contribution to journalArticle

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Abstract

CWR22 has been a valuable xenograft model for the study of prostate cancer progression from an androgen-dependent tumor to one that grows in castrated animals. Herein, we report the identification and characterization of a novel androgen receptor (AR) mutation occurring in a relapsed tumor (CWR22R-2152) and in the CWR22Rv1 cell line established from it. The mutation was not detected in the original, hormone-dependent CWR22 xenograft, indicating that this change occurred during the progression to androgen independence. It is characterized by an in-frame tandem duplication of exon 3 that encodes the second zinc finger of the AR DNA-binding domain. Accordingly, immunoblot analyses demonstrated the expression of an AR species having an approximately 5-kDa increase in size relative to the LNCaP AR. This was accompanied by a COOH-terminally truncated AR species migrating with a relative mass of 75-80 kDa, referred to as ARΔLBD because it lacks the ligand-binding domain. By recreating the exon 3 duplication mutation in a wild-type AR expression construct, the generation of ARΔLBD could be recapitulated. Whereas ARΔLBD exhibited constitutive nuclear localization and DNA binding, these functions in the full-length AR remained androgen dependent. The CWR22Rv1 AR repertoire displayed dose-dependent, androgen-responsive transcriptional transactivation in reporter assays, albeit to a lesser extent in comparison with LNCaP. This cell line also expressed low levels of prostate-specific antigen mRNA and did not express or secrete detectable levels of prostate-specific antigen protein in androgen-depleted medium or in response to physiological androgenic stimulation. In summary, the CWR22Rv1 cell line displays both androgen-responsive and androgen-insensitive features due, at least in part, to a novel insertional mutation of the AR.

Original languageEnglish (US)
Pages (from-to)6606-6614
Number of pages9
JournalCancer Research
Volume62
Issue number22
StatePublished - Nov 15 2002

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Androgen Receptors
Heterografts
Prostatic Neoplasms
Androgens
Cell Line
Mutation
Prostate-Specific Antigen
Exons
DNA
Zinc Fingers
Transcriptional Activation
Neoplasms
Hormones
Ligands
Messenger RNA

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Characterization of a novel androgen receptor mutation in a relapsed CWR22 prostate cancer xenograft and cell line. / Tepper, Clifford G; Boucher, David L.; Ryan, Philip E.; Ma, Ai Hong; Xia, Liang; Lee, Li Fen; Pretlow, Thomas G.; Kung, Hsing-Jien.

In: Cancer Research, Vol. 62, No. 22, 15.11.2002, p. 6606-6614.

Research output: Contribution to journalArticle

Tepper, CG, Boucher, DL, Ryan, PE, Ma, AH, Xia, L, Lee, LF, Pretlow, TG & Kung, H-J 2002, 'Characterization of a novel androgen receptor mutation in a relapsed CWR22 prostate cancer xenograft and cell line', Cancer Research, vol. 62, no. 22, pp. 6606-6614.
Tepper, Clifford G ; Boucher, David L. ; Ryan, Philip E. ; Ma, Ai Hong ; Xia, Liang ; Lee, Li Fen ; Pretlow, Thomas G. ; Kung, Hsing-Jien. / Characterization of a novel androgen receptor mutation in a relapsed CWR22 prostate cancer xenograft and cell line. In: Cancer Research. 2002 ; Vol. 62, No. 22. pp. 6606-6614.
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abstract = "CWR22 has been a valuable xenograft model for the study of prostate cancer progression from an androgen-dependent tumor to one that grows in castrated animals. Herein, we report the identification and characterization of a novel androgen receptor (AR) mutation occurring in a relapsed tumor (CWR22R-2152) and in the CWR22Rv1 cell line established from it. The mutation was not detected in the original, hormone-dependent CWR22 xenograft, indicating that this change occurred during the progression to androgen independence. It is characterized by an in-frame tandem duplication of exon 3 that encodes the second zinc finger of the AR DNA-binding domain. Accordingly, immunoblot analyses demonstrated the expression of an AR species having an approximately 5-kDa increase in size relative to the LNCaP AR. This was accompanied by a COOH-terminally truncated AR species migrating with a relative mass of 75-80 kDa, referred to as ARΔLBD because it lacks the ligand-binding domain. By recreating the exon 3 duplication mutation in a wild-type AR expression construct, the generation of ARΔLBD could be recapitulated. Whereas ARΔLBD exhibited constitutive nuclear localization and DNA binding, these functions in the full-length AR remained androgen dependent. The CWR22Rv1 AR repertoire displayed dose-dependent, androgen-responsive transcriptional transactivation in reporter assays, albeit to a lesser extent in comparison with LNCaP. This cell line also expressed low levels of prostate-specific antigen mRNA and did not express or secrete detectable levels of prostate-specific antigen protein in androgen-depleted medium or in response to physiological androgenic stimulation. In summary, the CWR22Rv1 cell line displays both androgen-responsive and androgen-insensitive features due, at least in part, to a novel insertional mutation of the AR.",
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AU - Pretlow, Thomas G.

AU - Kung, Hsing-Jien

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AB - CWR22 has been a valuable xenograft model for the study of prostate cancer progression from an androgen-dependent tumor to one that grows in castrated animals. Herein, we report the identification and characterization of a novel androgen receptor (AR) mutation occurring in a relapsed tumor (CWR22R-2152) and in the CWR22Rv1 cell line established from it. The mutation was not detected in the original, hormone-dependent CWR22 xenograft, indicating that this change occurred during the progression to androgen independence. It is characterized by an in-frame tandem duplication of exon 3 that encodes the second zinc finger of the AR DNA-binding domain. Accordingly, immunoblot analyses demonstrated the expression of an AR species having an approximately 5-kDa increase in size relative to the LNCaP AR. This was accompanied by a COOH-terminally truncated AR species migrating with a relative mass of 75-80 kDa, referred to as ARΔLBD because it lacks the ligand-binding domain. By recreating the exon 3 duplication mutation in a wild-type AR expression construct, the generation of ARΔLBD could be recapitulated. Whereas ARΔLBD exhibited constitutive nuclear localization and DNA binding, these functions in the full-length AR remained androgen dependent. The CWR22Rv1 AR repertoire displayed dose-dependent, androgen-responsive transcriptional transactivation in reporter assays, albeit to a lesser extent in comparison with LNCaP. This cell line also expressed low levels of prostate-specific antigen mRNA and did not express or secrete detectable levels of prostate-specific antigen protein in androgen-depleted medium or in response to physiological androgenic stimulation. In summary, the CWR22Rv1 cell line displays both androgen-responsive and androgen-insensitive features due, at least in part, to a novel insertional mutation of the AR.

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